Field of the Invention
The present invention is generally directed toward methods of characterizing nanoparticles, particularly nanoparticle complexes comprising an organic species such as a biomolecule or pharmaceutical compound. In certain embodiments, the present invention utilizes two-dimensional fluorescence difference spectroscopy in order to generate a unique identifier or “fingerprint” for the particular nanoparticle or nanoparticle complex that can be used to characterize the nanoparticle or nanoparticle complex.
Description of the Prior Art
In the last 10-15 years, nanotechnology has emerged as an area of great interest in the fields of physics and chemistry. While great numbers of various types of nanoparticles have been synthesized, including elemental nanoparticles and nanoparticle compounds and composites, and the physiochemical properties of these nanoparticles are fairly well understood, there exists a fundamental, if not disturbing, gap regarding the understanding of the biochemical and biological activities of these nanomaterials. In particular, there is a lack of understanding of the effect that many nanomaterials have on the structure-function of nucleic acids and proteins, cell metabolism, and cell signaling.
Due to this lack of understanding, many areas of nanomaterial research stand at an impasse. Materials originally thought to be inert are not. For example, despite the fact that earth's organisms are carbon-based, there is an alarming and steadily growing literature that suggests that carbon nanotubes (CNTs) can be quite toxic to cells and tissues. This seems to shadow, if not plague, many of the first generation nanomaterials that were studied including, but not limited to, silicon dioxide, quantum dots and others.
Nanoparticles have been proposed as platforms for delivery of various therapeutic materials, such as proteins and nucleic acids, into a human or animal body or as biosensors. In order to effectively design nanomaterials for these and other applications, it is necessary to develop a quantitative understanding of how nanoparticles perform and behave in the biological environment. More specifically, to study the important nanobio interface, it is necessary to have a means of quantifying the nanoparticle surface in such a way as biomolecular interaction could be directly detected.
Current methods of characterizing the structure of nanoparticle complexes include electron microscopy, dynamic laser light scatter, zeta potential analysis, and traditional fluorescence and absorbance spectral analyses, which require multiple techniques and data to piece together the information needed. Thus, these current methods of characterizing nanoparticle complexes are quite time intensive and may be insufficient to adequately characterize the particles. Accordingly, there is a need for methods of accurately characterizing nanoparticles and their biomolecular complexes more quickly, more simply, and quantitatively.